Agent For Growing Fibroblasts

ABSTRACT

An agent for growing fibroblasts contains a yeast extract and a safflower extract as effective constituents. A method of growing fibroblasts comprises using a yeast extract and a safflower extract. An anti-aging agent for skin contains a yeast extract and a safflower extract as effective constituents.

TECHNICAL FIELD

This invention relates to an agent for growing fibroblasts. Thisinvention particularly relates to an agent for growing fibroblastscapable of promoting the growth of aged fibroblasts. This invention alsorelates to an external preparation for skin, which preparation containsthe agent for growing fibroblasts.

BACKGROUND ART

Recently, keeping the healthy and beautiful skins has become a matter ofgreat concern without distinction of age or sex. The skin is principallyclassified into three layers, i.e., the epidermis, the derma, and thehypodermic tissue. Of the three layers, the derma is markedly importantfor keeping the skin structure. The derma is a binding tissue having astructure in which, principally, fibroblasts, collagen fibers, elasticfibers (elastin), and the like, spread in a composite, three-dimensionalpattern. The proteins, such as collagen, which constitute the fibers,are produced by the fibroblasts. The fibroblasts keep the balance ofsynthesis and decomposition of the proteins and keep the condition ofthe binding tissue. The strength, the stretchability, and the elasticityof the skin are thereby maintained.

Due to internal factors, such as aging, and external factors, such asultraviolet light and active oxygen, the binding tissue of the dermadecreases, and the functions which the skin keeps originally, such ascontractibility, flexibility, and moisturizing character, become weak.As a result, various troubles of the skin, such as the occurrence ofwrinkles and sagging, and the lowering of suppleness and elasticity,occur. It is considered that the important factors for the skin agingsymptoms as described above are the decrease of the number of thefibroblasts of the derma of the skin, the lowering of the functions ofthe fibroblasts of the derma of the skin, and the decrease of matrixfibers which are produced by the fibroblasts. Therefore, in order for ananti-aging external preparation for skin having an action of preventingand improving skin aging to be obtained, attempts have heretofore beenmade to search and blend a constituent having a function of promotingthe growth of fibroblasts.

For example, in cited literatures 1 and 2, it is reported that extractsof plants, such as Lens esculenta, Piper longum, Centella asiatica,Ocimum sanctum, Ocimum tenuiflorum, and Ocimum album, and a hibiscusextract have the fibroblast growth activity, and that the substancesdescribed above may be blended in anti-aging cosmetic preparations, andthe like.

However, in the cases of the skin aging, cellular proliferativepotential of fibroblast decreasing with age, such as the lowering of acell division rate of the fibroblasts, occurs. With the conventionalagents for growing fibroblasts, the problems are encountered in that,though the conventional agents for growing fibroblasts are effective foryoung fibroblasts, the conventional agents for growing fibroblasts arenot always capable of exhibiting a sufficient growth promoting effectwith respect to aged fibroblasts. Thus there is a strong demand fordevelopment of a novel substance which is capable of enhancing thegrowth of the aged fibroblasts.

It has been known that a yeast extract has a collagen productionpromoting action (in patent literature 3), a moisturizing effect (inpatent literature 4), a cell activating action, and a hyaluronic acidproduction promoting action (in patent literature 5). The yeast extractis blended in external preparations for skin, such as the cosmeticpreparations.

In the field of Chinese medicine, it has been known that a safflowerextract has a vitiated blood improving action. For the purposes ofbeauty culture, the safflower extract is used for improvement of thecold of the whole body (improvement of blood flow at skin distal areas),the improvement of a pimple, and treatment for atopic dermatitis (inpatent literature 6). Also, it has been reported that the safflowerextract has elastase inhibitory activity (in patent literature 7).Further, the safflower extract is blended in an anti-aging externalpreparation for skin, or the like, for obtaining a cell activatingaction (in patent literature 8).

RELATED ART LITERATURES

[Patent literature 1]

Japanese Unexamined Patent Publication No. 2008-184441

[Patent literature 2]

Japanese Patent No. 3748941

[Patent literature 3]

International Patent Publication No. WO 2004/075621

[Patent literature 4]

Japanese Patent Publication No. 6 (1994)-23088

[Patent literature 5]

Japanese Unexamined Patent Publication No. 8 (1996)-163983

[Patent literature 6]

Japanese Patent No. 3839438

[Patent literature 7]

Japanese Unexamined Patent Publication No. 2000-319189

[Patent literature 8]

Japanese Patent No. 3641699

The primary object of the present invention is to provide an agent forgrowing fibroblasts, which is capable of enhancing growth of not onlyyoung fibroblasts but also aged fibroblasts, and which is safe andeffective.

DISCLOSURE OF THE INVENTION

The inventors have found that, in cases where a yeast extract and asafflower extract are used in combination, cell growth is capable ofbeing promoted markedly even with respect to aged fibroblasts. Thepresent invention is based upon the findings described above. Forexample, as described in patent literature 8 cited above, each of theyeast extract and the safflower extract has heretofore been known as acell activating agent. However, nothing has heretofore been reported asto the effect on the fibroblasts, the cellular proliferative potentialof which has become decreased with age. Unexpectedly, in cases where thetwo constituents described above are used in combination, the growth ofthe aged fibroblasts is capable of being enhanced synergistically.

The present invention provides an agent for growing fibroblasts,containing a yeast extract and a safflower extract as effectiveconstituents.

The present invention also provides a method of growing fibroblasts,wherein a yeast extract and a safflower extract are used.

The present invention further provides an anti-aging agent for skin,containing a yeast extract and a safflower extract as effectiveconstituents.

In cases where the yeast extract or the safflower extract is used alone,a sufficient effect of promoting the growth is not obtained with respectto the aged fibroblasts. However, in cases where the two constituentsdescribed above are used in combination, a large effect of promoting thecell growth is obtained with respect to the aged fibroblasts as well aswith respect to the young fibroblasts. Therefore, the skin aging isprevented effectively.

The term “aged fibroblasts” as used herein means the fibroblasts whichhave suffered from the decreasing in functions, such as the cellularproliferative potential and the extracellular matrix producingcapability, with age or due to adverse effects of ultraviolet light,active oxygen, and the like.

The yeast extract should preferably be the yeast extract having beenprepared from the yeast, which has been cultured in a nutrient culturemedium containing glycosaminoglycan and has been subjected toultraviolet light irradiation processing and/or hydrogen peroxideprocessing. It has been found that, in cases where the yeast is culturedin the glycosaminoglycan-containing nutrient culture medium in thepresence of a stress due to ultraviolet light and/or hydrogen peroxide,a cell protecting constituent is produced. By use of the yeast extracthaving been prepared from the cultured yeast as described above, thegrowth of the fibroblasts is enhanced more effectively.

The present invention still further provides an external preparation forskin, containing the agent for growing fibroblasts in accordance withthe present invention. For example, in cases where the agent for growingfibroblasts in accordance with the present invention is blended in theexternal preparation for skin, such as a cosmetic preparation, it ispossible to prevent or improve the decrease of the function and numberof the fibroblasts in the binding tissue of the derma with age, and thelike, and to effectively prevent or improve the symptoms of the skinaging, such as the occurrence of the wrinkles and the sagging, and thelowering of the suppleness and the elasticity.

In accordance with the present invention, wherein the yeast extract andthe safflower extract are used in combination, the large effect ofpromoting the cell growth is obtained with respect to not only the youngfibroblasts but also the aged fibroblasts. Also, the fibroblasts in thederma of the aged skin are multiplied, and the suppleness and theelasticity of the skin are enhanced in a safe and effective manner, suchthat no adverse effect may occur on the skin. Therefore, by theapplication of the agent for growing fibroblasts in accordance with thepresent invention or the external preparation for skin, which containsthe agent for growing fibroblasts in accordance with the presentinvention, to the skin, the symptoms of the skin aging, such as theoccurrence of wrinkles and sagging, and the lowering of the supplenessand the elasticity, are prevented or improved effectively.

BRIEF DESCRIPTION OF THE DRAWING

FIG. 1 is a graph showing an effect of a combination of a safflowerextract and a yeast extract upon growth of young or aged fibroblasts.

BEST MODE FOR CARRYING OUT THE INVENTION

The agent for growing fibroblasts, or the like, in accordance with thepresent invention contains the yeast extract and the safflower extractas the effective constituents.

As the yeast extract, it is possible to employ an extract of the yeastwith a polar solvent; a yeast decomposition product, which results frombacteriolysis of the yeast with self-digestion, acidolysis, enzymatichydrolysis, or the like, and which has been subjected to filtration; oran extract obtained from processing, in which the yeast decompositionproduct as described above is dried and then subjected to extractionwith a polar solvent, or the like.

As a fungus body for the preparation of the yeast extract, it ispossible to employ preferably an Ascomycetes yeast, e.g., a yeast of agenus belonging to an Endomycetaceae family, such as an Eremascus genusor an Endomyces genus; a yeast of a genus belonging to aSaccharomycetaceae family, such as a Schizosaccharomyces genus, aNadsonia genus, a Saccharomycodes genus, a Hanseniaspora genus, aWickerhamia genus, a Saccharomyces genus, a Kluyveromyces genus, aLodderomyces genus, a Wingea genus, an Endomycopsis genus, a Pichiagenus, a Hansenula genus, a Pachysolen genus, a Citeromyces genus, aDebaryomyces genus, a Schwanniomyces genus, a Dekkera genus, aSaccharomycopsis genus, or a Liporayces genus; or a yeast of a genusbelonging to a Spermophthoraceae family, such as a Spermophthora genus,an Eremothecium genus, a Crebrothecium genus, an Ashbya genus, aNematospora genus, a Metschnikowia genus, or a Coccidiasous genus.Particularly, from the view point of availability, and the like, theyeast extract should preferably be prepared from a fungus body, such asthe yeast of the Saccharomyces genus, e.g., a beer yeast, a sake yeast,or a baker's yeast. In the present invention, it is also possible toemploy a yeast extract commercially available under the trade name of“Yeast Extract.”

As a solvent for the extraction, it is possible to employ water;physiological saline; a phosphate buffer; phosphate buffered saline; apolar organic solvent, e.g., a lower alcohol, such as methanol, ethanol,propanol, or isopropanol; a polyhydric alcohol, such as 1,3-butyleneglycol, propylene glycol, dipropylene glycol, or glycerol; an ether,such as diethyl ether or dipropyl ether; an ester, such as ethyl acetateor butyl acetate; or a ketone, such as acetone or ethyl methyl ketone.One kind of the solvent or at least two kinds of the solvents selectedfrom the above-enumerated solvents are used. In cases where solventextraction is performed, the yeast or the yeast decomposition productmay be subjected to freeze-drying and/or pulverizing and may then besubjected to the extraction. Also, homogenizing may be performed in theextraction solvent, and ultrasonic crushing may be performed. Theextraction temperature ordinarily falls within the range ofapproximately 0° C. to a temperature equal to at most the boilingtemperature of the solvent. Also, the extraction time varies inaccordance with the kind of the extraction solvent and the extractiontemperature and ordinarily falls within the range of approximately onehour to approximately five days.

In the present invention, as the yeast extract, it is possible to usepreferably the yeast extract having been prepared from the yeast, whichhas been cultured in the nutrient culture medium containingglycosaminoglycan and has been subjected to the ultraviolet lightirradiation processing and/or the hydrogen peroxide processing. It hasbeen found that, in cases where the yeast is cultured in the presence ofa stress due to ultraviolet light and/or hydrogen peroxide, the cellprotecting constituent, which protects the cells from the stress, isproduced. Further, in such cases, by the addition of nutrient peptone,glycosaminoglycan, or the like, to the culture medium, the response tothe stress is enhanced. A process for producing the yeast extract, orthe like, is described in, for example, U.S. Pat. No. 6,461,857. Forexample, the yeast extract may be prepared with a process, wherein aSaccharomyces cerevisiae yeast, which is one kind of the baker's yeast,is cultured in a nutrient culture medium having been added withnon-animal derived glycosaminoglycan, wherein a stress is loaded to thecultured yeast by the addition of hydrogen peroxide in a sublethalproportion, e.g. in a proportion falling within the range ofapproximately 0.1% by mass to 2% by mass with respect to the total massof the cultured yeast, and optionally by the irradiation of a sublethaldose of the ultraviolet light (e.g., UVA/UVB irradiation with anintensity of 31.5 mJ/cm²), and wherein the cultured yeast having thusbeen obtained is solubilized by self-digestion, acidolysis, or the like,and is then subjected to drying, extraction with water, filtration, andthe like. Also, a yeast extract having been prepared from a yeast, whichhas been cultured in a culture medium containing non-animal derivedglycosaminoglycan and has been subjected to the ultraviolet lightirradiation processing and the hydrogen peroxide processing, iscommercially supplied under the trade name “Biodyne EMPP (trademark)” byArch Personal Care Products L. P.

The safflower extract may be prepared by performing solvent extractionof a safflower (Carthamus tinctorius L.). It is possible to use each ofparts of the safflower, such as a flower, a leaf, a stalk, a root stalk,and a root, or the whole of the safflower. It is preferable to use theflower of the safflower used as a crude drug “Kouka (Carthami Flos.)”

The safflower may be subjected to the extracting operation in a crudestate. However, from the view point of the extraction efficiency, theextraction should preferably be performed after the processing, such asslicing, drying, or pulverizing, has been performed. The extraction isperformed by dipping in the extraction solvent. In order for theextraction efficiency to be enhanced, stirring may be performed, or thehomogenizing may be performed in the extraction solvent. The extractiontemperature ordinarily falls within the range of approximately 5° C. toa temperature equal to at most the boiling temperature of the solvent.Also, the extraction time ordinarily falls within the range ofapproximately four hours to approximately 14 days.

As the extraction solvent, it is possible to employ water; physiologicalsaline; a phosphate buffer; phosphate buffered saline; a polar organicsolvent, e.g., a lower alcohol, such as methanol, ethanol, propanol,isopropanol, or butanol; a polyhydric alcohol, such as 1,3-butyleneglycol, propylene glycol, dipropylene glycol, 1, 2-pentanediol, orglycerol; an ether, such as ethyl ether or propyl ether; an ester, suchas ethyl acetate or butyl acetate; or a ketone, such as acetone or ethylmethyl ketone; or a mixed solvent containing at least two kinds of thesolvents selected from the above-enumerated solvents.

For example, in the present invention, it is possible to use preferablya safflower extract having been obtained from the processing wherein theflower of the safflower is subjected to the extraction with 50%1,3-butylene glycol and the filtration. Also, an extract commerciallyavailable as the safflower extract may be used in the present invention.

As the agent for growing fibroblasts, or the like, in accordance withthe present invention, the yeast extract and the safflower extract maybe employed directly. Alternatively, the agent for growing fibroblasts,or the like, in accordance with the present invention may be used in anarbitrary form, such as a solution, a suspension, an emulsion, a creamagent, an ointment agent, a gel agent, or a powdered medicine, by theaddition of an arbitrary appropriate auxiliary, such as a carrier, adiluting agent, a stabilizing agent, a buffer agent, a pH adjustingagent, or a solvent. Also, the agent for growing fibroblasts, or thelike, in accordance with the present invention may be encapsulated in aliposome, a micro-capsule, or the like. Further, besides the essentialconstituents described above, the agent for growing fibroblasts, or thelike, in accordance with the present invention may be added with astabilizing agent and an absorption promoting agent, such as ananti-oxidizing agent, an antiseptic agent, or a ultraviolet lightabsorbing agent, such that the effects of the present invention may notbe affected adversely.

The agent for growing fibroblasts, or the like, in accordance with thepresent invention may be contained in an external preparation base, orthe like, and the external preparation for skin may thereby be prepared.

In so far as the effects of the present invention are accomplished, nolimitation is imposed upon the blending proportion of each of the yeastextract and the safflower extract in the external preparation for skinin accordance with the present invention. However, the blendingproportion of the yeast extract with respect to the total mass of theexternal preparation should preferably fall within the range of 0.00001%by mass to 10% by mass, should more preferably fall within the range of0.001% by mass to 5% by mass, and should most preferably fall within therange of 0.005% by mass to 1% by mass. Also, the blending proportion ofthe safflower extract with respect to the total mass of the externalpreparation should preferably fall within the range of 0.00001% by massto 10% by mass, should more preferably fall within the range of 0.001%by mass to 5% by mass, and should most preferably fall within the rangeof 0.01% by mass to 1% by mass.

The external preparation for skin in accordance with the presentinvention may embrace cosmetic preparations, pharmaceuticalpreparations, quasi-drugs, and the like, and may more preferably beapplied broadly to the cosmetic preparations. Also, the externalpreparation for skin in accordance with the present invention may takeon an arbitrary preparation form, such as an aqueous solution type, asolubilized type, an emulsion type, an oily liquid type, a gel type, apaste type, an ointment type, an aerosol type, a water-oil two layertype or a water-oil-powder three layer type. Further, the externalpreparation for skin in accordance with the present invention may takeon a form carried by a sheet-shaped base material.

Furthermore, the external preparation for skin in accordance with thepresent invention may take on an arbitrary product form and may be usedfor an arbitrary use application, such that the effects of the presentinvention may be achieved. For example, the external preparation forskin in accordance with the present invention may be applied to basicskin care cosmetics, such as a cleansing preparation, a cosmetic lotion,a cosmetic base lotion or cream, a milky lotion, a cream, a gel, anessence (a beauty liquid), a pack, or a mask; a make-up cosmeticpreparation, such as a foundation; a sunscreen cosmetic preparation,such as a sunscreen lotion or cream; a body cosmetic preparation, suchas a hand lotion or cream, a leg lotion or cream, or a body lotion orcream; or a toiletry product, such as a body soap or a toilet soap.

If necessary, besides the essential constituents described above, theexternal preparation for skin in accordance with the present inventionmay be blended appropriately with a different arbitrary constituentordinarily used in external preparations for skin, such as cosmeticpreparations and pharmaceutical preparations, and may be produced by aconventional procedure in accordance with the desired preparation form.For example, if necessary, the external preparation for skin inaccordance with the present invention may be blended appropriately witha whitening agent, a moisturizing agent, an anti-oxidizing agent, anoily constituent, an ultraviolet light absorbing agent, a surface-activeagent, a thickening agent, an alcohol, a powder constituent, a coloringagent, an aqueous constituent, water, or a skin nutrient agent. Also, itis possible to blend appropriately a sequestering agent, such as anedetic acid disodium salt, an edetic acid trisodium salt, a sodiumcitrate, sodium polyphosphate, sodium metaphosphate, or gluconic acid; amedicine, such as caffeine, tannin, verapamil, tranexamic acid or aderivative thereof, a licorice extract, glabridin, a hot water extractof a fruit of narrow leaf-firethorn (Pyracantha angustifolia Schneid.),a crude drug, tocopheryl acetate, glycyrrhizinic acid or a derivativethereof or a salt thereof; a whitening agent, such as vitamin C,magnesium ascorbyl phosphate, ascorbyl glucoside, arbutin, or kojicacid; or a saccharide, such as glucose, fructose, mannose, sucrose, ortrehalose.

EXAMPLES

The present invention will further be illustrated by the followingnon-limitative examples. In the examples described below, Biodyne EMPP(trademark) (supplied by Arch Personal Care Products L. P.) was employedas the yeast extract, and Pharcolex Safflower B (supplied by ICHIMARUPHARCOS Co., Ltd.) was employed as the safflower extract.

Study of Effect of Promoting Growth of Fibroblasts

Fibroblasts having been derived from the human newborn foreskin wereused as “young fibroblasts”, and fibroblasts having been derived fromthe derma of the skin of a septuagenarian female were used as “agedfibroblasts”.

As for each of the young fibroblasts and the aged fibroblasts, thefibroblasts were cultured in a 10% FBS-containing DIEM culture mediumfor 24 hours. Thereafter, the culture medium was replaced by a 0.5%FBS-containing DMEM culture medium, which had been added with 0.1% of asafflower extract, a 0.5% FBS-containing DMEM culture medium, which hadbeen added with 0.01% of a yeast extract, or a 0.5% FBS-containing DMEMculture medium, which had been added with 0.1% of the safflower extractand 0.01% of the yeast extract, and the fibroblasts were cultured stillfurther for 48 hours. Also, as a control, the culture processing wasperformed in the same manner in a 0.5% FBS-containing DMEM culturemedium, which had not been added with the test substance. At the time ofthe end of the culture processing, respiration activity of thefibroblasts was measured by use of an Alamar Blue reagent and was usedas an index for the growth of the fibroblasts. A relative fibroblastgrowth (%) was calculated with respect to the respiration activity ofthe control, which was taken as 100%.

The results illustrated in FIG. 1 were obtained. With the safflowerextract alone, as for both of the young fibroblasts and the agedfibroblasts, the growth of the fibroblasts was enhanced slightly, andthe effect of promoting the growth of the fibroblasts was small. Also,with the yeast extract alone, the growth of the young fibroblasts wasenhanced markedly, but the effect of promoting the growth of thefibroblasts with respect to the aged fibroblasts was small. However, bythe addition of the safflower extract and the yeast extract incombination, the effect of promoting the growth of the fibroblasts,particularly with respect to the aged fibroblasts, was enhancedsynergistically, and the growth of the aged fibroblasts was enhanced toa level equivalent to the level of the growth of the young fibroblasts.

It was suggested that, by the use of the safflower extract and the yeastextract in combination, the growth of the aged fibroblasts, the cellularproliferative potential of which had lowered with age, was enhancedmarkedly, the growth of the fibroblasts was enhanced even in the casesof the aged skin, and the symptoms of the skin aging, such as theoccurrence of wrinkles and sagged skin, and the lowering of thesuppleness and the elasticity, were prevented or improved effectively.

Examples for the external preparation for skin containing the agent forgrowing fibroblasts in accordance with the present invention will beillustrated hereinbelow. The blending proportions were expressed interms of % by mass with respect to the total mass of the composition.

Example 1 W/O Cream

(% by mass) (1) Dimethylpolysiloxane 3.0 (2)Decamethylcyclopentasiloxane 13.0 (3) Dodecamethylcyclohexasiloxane 5.0(4) Polyoxyethylene-methylpolysiloxane 1.0 copolymer (5)3-Tris(trimethylsiloxy)silyl- 1.0 propylcarbamic acid pullulan (6)Decamethylcyclopentasiloxane blend of 5.0 crosslinked typepolymethylsiloxane (Dow Corning 9040 Silicone Elastomer Blend TM,supplied by Dow Corning Co., Ltd.) (7) Retinol acetate 0.1 (8)Para-hydroxybenzoic acid ester Appropriate quantity (9) L-MentholAppropriate quantity (10) Trimethylsiloxysilicic acid 2.0 (11) Ethanol2.0 (12) Glycerol 3.0 (13) Dipropylene glycol 5.0 (14) PolyethyleneGlycol 6000 5.0 (15) Sodium hexametaphosphate 0.05 (16) Tocopherolacetate 0.1 (17) Caffeine 0.1 (18) Fennel extract 0.1 (19) Hamamelisextract 0.1 (20) Safflower extract 1.0 (21) Yeast extract 1.0 (22)Edetic acid trisodium salt 0.05 (23) Dimorpholinopyridazinone 0.01 (24)Trimethoxycinnamic acid 0.1 methylbis(trimethylsiloxy) silylisopentyl(25) Inorganic composite particles 5.0 (Cover Leaf AR-80TM, supplied byJGC Catalysts and Chemicals Ltd.) (26) Yellow iron oxide Appropriatequantity (27) Cobalt titanate Appropriate quantity (28)Dimethyldistearylammonium hectorite 1.5 (29) Polyvinyl alcohol 0.1 (30)Hydroxyethyl cellulose 0.1 (31) Sodium acrylate/2-acrylamido-2- 0.1methylpropanesulfonic acid copolymer (SIMULGEL EGTM, supplied by SEPPICCo., Ltd.) (32) (Acryloyldimethyltaurineammonium/VP) 0.1 copolymer(Aristoflex AVCTM, supplied by Clariant Co., Ltd.) (33) PerfumeAppropriate quantity (34) Deionized water Remainder

Example 2 Milky Lotion

(% by mass) Petrolatum 5 Behenyl alcohol 0.5 Batyl alcohol 0.5 Glycerol7 1,3-Butylene glycol 7 1,2-Pentanediol 1 Xylitol 3 Polyethylene Glycol20000 2 Hydrogenated oil 2 Jojoba oil 2 Squalane 5 Isostearic acid 0.5Tetra-2-ethylhexanoic acid pentaerythritol 2 Polyoxyethylenehydrogenated caster oil 0.5 Lauryldimethylaminoacetic acid betaine 0.4Potassium hydroxide Appropriate quantity Sodium pyrosulfite 0.01 Sodiumhexametaphosphate 0.05 Dipotassium glycyrrhizinate 0.05 Trimethylglycine3 Arbutin 3 Yeast extract 0.1 Tocopherol acetate 0.1 Thiotaurine 0.1Safflower extract 1 Sophora flavescens extract 0.1 Red iron oxideAppropriate quantity Quince seed extract 0.1 Carboxyvinyl polymer 0.2Phenoxy ethanol Appropriate quantity Deionized water Remainder

Example 3 Milky Lotion

(% by mass) Dimethylpolysiloxane 3 Decamethylcyclopentasiloxane 4Ethanol 5 Glycerol 6 1,3-Butylene glycol 5Polyoxyethylenemethylglucoside 3 Sunflower oil 1 Squalane 2 Potassiumhydroxide 0.1 Sodium hexametaphosphate 0.05 Hydroxypropyl-β-cyclodextrin0.1 Dipotassium glycyrrhizinate 0.05 Loquat leaf extract 0.1 Monosodiumglutamate 0.05 Fennel extract 0.1 Yeast extract 0.1 Lavender oil 0.1Safflower extract 0.1 Rehmannia Chinensis Root Extract 0.1Dimorpholinopyridazinone 0.1 Xanthan gum 0.1 Carboxyvinyl polymer 0.1Acrylic acid-alkyl methacrylate copolymer 0.1 (PEMULEN TR-1) Red ironoxide Appropriate quantity Yellow iron oxide Appropriate quantityParaben Appropriate quantity Deionized water Remainder

The external preparation for skin of each of the examples describedabove exhibited the excellent effect of improving wrinkles by beingapplied to the aged skin.

1-9. (canceled)
 10. An agent for growing fibroblasts, containing a yeastextract and a safflower extract as effective constituents.
 11. An agentfor growing fibroblasts as defined in claim 10, wherein the yeastextract is a yeast extract having been prepared from a yeast, which hasbeen cultured in a nutrient culture medium containing glycosaminoglycanand has been subjected to ultraviolet light irradiation processingand/or hydrogen peroxide processing.
 12. An agent for growingfibroblasts as defined in claim 10, wherein the fibroblasts are agefibroblasts.
 13. An agent for growing fibroblasts as defined in claim11, wherein the Fibroblasts are age fibroblasts.
 14. An externalpreparation for skin, containing an agent for growing fibroblasts asdefined in claim
 10. 15. An external preparation for skin as defined inclaim 14, wherein the external preparation for skin is a cosmeticpreparation.
 16. An external preparation for skin, containing an agentfor growing fibroblasts as defined in claim
 11. 17. An externalpreparation for skin as defined in claim 16, wherein the externalpreparation for skin is a cosmetic preparation.
 18. A method of growingfibroblasts, comprising using a yeast extract and a safflower extract.19. A method as defined in claim 18, wherein the yeast extract is ayeast extract having been prepared from a yeast, which has been culturedin a nutrient culture medium containing glycosaminoglycan and has beensubjected to ultraviolet light irradiation processing and/or hydrogenperoxide processing.
 20. A method as defined in claim 18, wherein thefibroblasts are age fibroblasts.
 21. A method as defined in claim 19,wherein the fibroblasts are age fibroblasts.
 22. An anti-aging agent forskin, containing a yeast extract and a safflower extract as effectiveconstituents.